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目的:采用代谢足迹技术探讨通天草提取物对Aβ损伤SAMP8小鼠原代海马神经元细胞的保护作用及其代谢机制。方法:采用MTT法测定细胞增值率测定Aβ损伤SAMP8小鼠原代海马神经元细胞的细胞增殖情况,在此基础上首次采用代谢足迹技术评价通天草提取物的疗效。聚焦关键代谢通路及相关代谢靶标,阐明其Aβ损伤SAMP8小鼠原代海马神经元细胞的发病机制和通天草提取物的作用机制。结果:MTT法测定细胞增值率结果 Aβ损伤SAMP8小鼠的原代海马神经元细胞的细胞活力明显减少,经代谢足迹技术研究发现,与同窝野生小鼠相比,Aβ损伤SAMP8小鼠的原代海马神经元细胞代谢异常主要集中在与神经细胞相关的叶酸代谢和牛磺酸代谢上,经高通量质谱解析及文献数据库检索确定了3个差异代谢物,分别是L-磺基丙氨酸(L-Cysteic acid)、二氢叶酸(Dihydrofolate)、分支酸(Chorismate),这些小分子代谢产物经通天草提取物干预后有明显的回调趋势。结论:通天草提取物对Aβ损伤SAMP8小鼠的原代海马神经元细胞具有一定程度的治疗作用,本次发现的3个生物标记物可能是Aβ损伤SAMP8小鼠的原代海马神经元细胞发病机制的潜在靶点,给予通天草提取物后这些标记物呈不同程度的回调趋势,为通天草提取物治疗阿尔兹海默病提供实验依据。
OBJECTIVE: To explore the protective effect and the metabolic mechanism of the extract ofTopia officinarum on primary cultured hippocampal neurons of SAMP8 mice with Aβtranscription injury. Methods: Cell proliferation rate was measured by MTT method. The primary cultured hippocampal neurons of SAMP8 mice were assayed by MTT assay. Focusing on the key metabolic pathways and related metabolic targets, elucidating the pathogenesis of Aβ-injured SAMP8 mouse primary hippocampal neurons and the mechanism of action of the extract of Tentia senticidus. Results: Cell proliferation rate was determined by MTT assay. The viability of primary hippocampal neurons in SAMP8 mice was significantly reduced by Aβ injury. The metabolic footprint analysis showed that compared with wild-type mice, Aβ-injured SAMP8 mice The metabolic abnormalities of hippocampal neurons were mainly concentrated on the folic acid metabolism and taurine metabolism related to nerve cells. Three different metabolites were identified by high-throughput mass spectrometry and literature search, which were L-cysteic acid L-Cysteic acid, Dihydrofolate and Chorismate. These small-molecule metabolites have a clear trend of callback after the intervention of these extracts. CONCLUSION: The extract ofTopia has a certain degree of therapeutic effect on primary hippocampal neurons damaged by Aβ in SAMP8 mice. The three biomarkers found in this study may be the primary hippocampal neuronal cells in Aβ-injured SAMP8 mice Mechanism of potential targets, given the extract of Viagra after these markers showed a trend of callback to varying degrees, to provide experimental evidence for the treatment of Alzheimer’s disease.